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目的观察原癌基因C-myc、H-ras及抑癌基因p16在硫酸铍致人胚肺成纤维细胞(HELF)损伤下的表达,探讨硫酸铍对离体培养HELF的影响。方法不同物质的量浓度硫酸铍(终物质的量浓度为2.0、20.0、100.0μmol/L)重复染毒HELF后,采用免疫组化SP法和甲基化特异性聚合酶链式反应(MSP)技术分别检测C-myc、H-ras蛋白及p16基因的表达。结果不同物质的量浓度染毒组培养至第12、24、36天时,C-myc蛋白的表达均明显高于对照组,差异有统计学意义(P<0.05,P<0.01),且具有剂量效应关系(r12=0.755,r24=0.822,r36=0.792,P<0.01);不同物质的量浓度染毒组在培养至第24、36天时H-ras蛋白表达明显高于对照组,差异有统计学意义(P<0.05或P<0.01),且具有剂量效应关系(r24=0.586,r36=0.845,P<0.01)。染毒组p16基因在培养至第24、36天时,第24天2.0μmol/L染毒组表达部分甲基化条带,其他均表达甲基化条带。结论硫酸铍在引起HELF损伤的情况下,可诱发HELFC-myc和H-ras蛋白异常表达、p16基因甲基化。
Objective To investigate the expression of protooncogene C-myc, H-ras and tumor suppressor gene p16 in human embryonic lung fibroblast (HELF) -induced injury induced by beryllium sulfate, and to explore the effect of beryllium sulfate on HELF in vitro. Methods The concentrations of different substances were determined by immunohistochemical SP method and methylation-specific polymerase chain reaction (MSP) after exposure to repeated exposure to beryllium sulfate (2.0,20.0,100.0μmol / L final concentration) Technology were detected C-myc, H-ras protein and p16 gene expression. Results The concentration of different substances in culture group reached the 12th, 24th and 36th day after exposure, the expression of C-myc protein was significantly higher than the control group, the difference was statistically significant (P <0.05, P <0.01) (R12 = 0.755, r24 = 0.822, r36 = 0.792, P <0.01). The levels of H-ras protein in the different concentration groups were significantly higher than those in the control group at 24 and 36 days (P <0.05 or P <0.01), and had dose-effect relationship (r24 = 0.586, r36 = 0.845, P <0.01). In the exposure group, the p16 gene was expressed on the 24th and 36th day, and the 2.0μmol / L exposure group on the 24th day showed some methylation bands, while others all expressed the methylation bands. Conclusion Beryllium sulphate can induce abnormal expression of HELFC-myc and H-ras protein and methylation of p16 gene in the condition of HELF injury.